DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log₁₀ GC/L (and up to 3.04 log₁₀ GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.     

Assessment and Evaluation of an Integrated Hybrid Anaerobic–Aerobic Sewage Treatment System for the Removal of Enteric Viruses

The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log₁₀ reductions of enteric viruses’ genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log₁₀ for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log₁₀ reduction which is very close to the overall log₁₀ reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log₁₀ reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.     

Metabolomic analysis of the selection response of Drosophila melanogaster to environmental stress: are there links to gene expression and phenotypic traits?

We investigated the global metabolite response to artificial selection for tolerance to stressful conditions such as cold, heat, starvation, and desiccation, and for longevity in Drosophila melanogaster. Our findings were compared to data from other levels of biological organization, including gene expression, physiological traits, and organismal stress tolerance phenotype. Overall, we found that selection for environmental stress tolerance changes the metabolomic ¹H NMR fingerprint largely in a similar manner independent of the trait selected for, indicating that experimental evolution led to a general stress selection response at the metabolomic level. Integrative analyses across data sets showed little similarity when general correlations between selection effects at the level of the metabolome and gene expression were compared. This is likely due to the fact that the changes caused by these selection regimes were rather mild and/or that the dominating determinants for gene expression and metabolite levels were different. However, expression of a number of genes was correlated with the metabolite data. Many of the identified genes were general stress response genes that are down-regulated in response to selection for some of the stresses in this study. Overall, the results illustrate that selection markedly alters the metabolite profile and that the coupling between different levels of biological organization indeed is present though not very strong for stress selection at this level. The results highlight the extreme complexity of environmental stress adaptation and the difficulty of extrapolating and interpreting responses across levels of biological organization.     

Seasonal changes in burrow geometry of the common mole rat (Rodentia: Bathyergidae)

Sociality in mole rats has been suggested to have evolved as a response to the widely dispersed food resources and the limited burrowing opportunities that result from sporadic rainfall events. In the most arid regions, individual foraging efficiency is reduced, and energetic constraints increase. In this study, we investigate seasonal differences in burrow architecture of the social Cryptomys hottentotus hottentotus in a mesic region. We describe burrow geometry in response to seasonal weather conditions for two seasons (wet and dry). Interactions occurred between seasons and colony size for the size of the burrow systems, but not the shape of the burrow systems. The fractal dimension values of the burrow systems did not differ between seasons. Thus, the burrow complexity was dependent upon the number of mole rats present in the social group.     

Bechstein’s bats maintain individual social links despite a complete reorganisation of their colony structure

Several social mammals, including elephants and some primates, whales and bats, live in multilevel societies that form temporary subgroups. Despite these fission–fusion dynamics, group members often maintain long-term bonds. However, it is unclear whether such individual links and the resulting stable social subunits continue to exist after a complete reorganisation of a society, e.g. following a population crash. Here, we employed a weighted network analysis on 7,109 individual roosting records collected over 4 years in a wild Bechstein’s bat colony. We show that, in response to a strong population decline, the colony’s two stable social subunits fused into a non-modular social network. Nevertheless, in the first year after the crash, long-term bonds were still detectable, suggesting that the bats remembered previous individual relationships. Our findings are important for understanding the flexibility of animal societies in the face of dramatic changes and for the conservation of social mammals with declining populations.

     

Warm forming die design,part I: Experimental validation of a novel thermal finite element modeling code

Finite element analysis (FEA) has become an invaluable tool in the design of sheet metal stamping dies and processes. FEA has gained widespread acceptance as the best method of optimizing dies for conventional stamping processes. More recently, FEA has been shown to be an effective method of designing tooling for sheet forming processes. In this work, an FEA based approach is applied to the warm stamping (warm forming) process. This work introduces a new thermal finite element analysis software called PASSAGE®/Forming (PASSAGE) that enables the up-front design of the thermal management of warm forming dies. This thermal finite element analysis software is designed to specifically handle the forming and optimization scenarios related to the heating of a stamping die while minimizing user interface time. In this work, PASSAGE has been applied to a simple block of steel embedded with cartridge heaters to validate the prediction capability of this software under two different heating conditions. The results show that PASSAGE is capable of predicting the actual steady-state temperature distribution within the block with an acceptable level of accuracy while yielding notable information to the user with respect to specifying power requirements. A finite element software package like PASSAGE is a valuable tool that will aid greatly in the implementation of warm forming as a manufacturing process beyond the scope of the laboratory and into production.     

Part based process performance monitoring (PbPPM)

The performances of completed manufacturing processes were evaluated using the surface response to excitation (SuRE) and Lamb wave methods. Both methods used the same piezoelectric elements attached to the surface of the workpiece. The SuRE method and the Lamb wave method were used to identify the structural changes created by welding, drilling, coating, filling a slot with glue, and composite patching. This study indicates that the tested methods are feasible for part based process performance monitoring (PbPPM) which evaluates the quality of the completed process with the sensors attached to the workpiece.     

Detection Method for Avian Influenza Viruses in Water

Recent events have shown that humans may become infected with some pathogenic avian influenza A viruses (AIV). Since soil and water, including lakes, rivers, and seashores, may be contaminated by AIV excreted by birds, effective methods are needed for monitoring water for emerging viruses. Combining water filtration with molecular methods such as PCR is a fast and effective way for detecting viruses. The objective of this study was to apply a convenient method for the detection of AIV in natural water samples. Distilled water and lake, river, and seawater were artificially contaminated with AIV (H5N3) and passed through a filter system. AIV was detected from filter membrane by real-time RT-PCR. The performance of Zetapor, SMWP, and Sartobind D5F membranes in recovering influenza viruses was first evaluated using contaminated distilled water. SWMP, which gave the highest virus recoveries, was then compared with a pre-filter combined GF/F filter membrane in a trial using natural water samples. In this study, the cellulose membrane SMWP was found to be practical for recovery of AIVs in water. Viral yields varied between 62.1 and 65.9% in distilled water and between 1 and 16.7% in natural water samples. The borosilicate glass membrane GF/F combined with pre-filter was also feasible in filtering natural water samples with viral yields from 1.98 to 7.33%. The methods described can be used for monitoring fresh and seawater samples for the presence of AIV and to determine the source of AIV transmission in an outbreak situation.     

Prevalence of Norovirus Infection in Children and Adults with Acute Gastroenteritis,Tehran, Iran, 2008–2009

Noroviruses are one of important agents that cause acute viral gastroenteritis worldwide. These viruses are belonging to Caliciviridae family and are genetically diverse. To date, there is no valuable data about prevalence of norovirus infection and the dominant genogroup/genotype among Iranian population. The objective of this study was to determine the frequency of norovirus infection in Iranian patients with gastroenteritis referred to three hospitals of Tehran and to specify the dominant genogroup/genotype of this virus among our study population. A total of 293 patients with acute gastroenteritis were included in the study. Detection of norovirus was performed using RT-PCR method and confirmed by direct sequencing with specific designed primers for capsid region of norovirus genome. Phylogenetic analysis was performed using the neighbor-joining method. Norovirus strains identified in our study were subsequently categorized according to previously defined genogroup/genotypes. Of these, norovirus GII was dominant genogroup. Sixty-five percent (17 of 26) of positive samples were determined as GII and 35% (9 of 26) were determined as GI, respectively, in 2008–2009. And among 8 sequenced strains of genogroup II the most frequent genotype was GII.3. The results of this study indicated that norovirus must be considered as one of the infectious causes of acute gastroenteritis among Iranian population. We also found that GII.3 is more prevalent in our study population. To the best of our knowledge there is limited data about the role of noroviruses in children and adults’ acute gastroenteritis among Iranian patients and this prevalence and genotyping report of norovirus infection could be remarkable for further studies.